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J Virol Methods. 2018 Oct;260:1-5. doi: 10.1016/j.jviromet.2018.06.020. Epub 2018 Jun 30.

Analysis of drug resistance mutations in whole blood DNA from HIV-1 infected patients by single genome and ultradeep sequencing analysis.

Author information

1
Department of Microbiology, Instituto de Investigación Hospital 12 de Octubre, Madrid, Spain.
2
HIV Unit, Department of Internal Medicine, Instituto de Investigación Hospital 12 de Octubre, Madrid, Spain.
3
Department of Microbiology, Instituto de Investigación Hospital 12 de Octubre, Madrid, Spain. Electronic address: rafael.delgado@salud.madrid.org.

Abstract

In HIV-1 infected patients blood CD4+ T lymphocytes could be a valuable target to analyse drug resistance mutations (DRM) selected over the course of the infection. However, detection of viral resistance mutations in cellular DNA by standard genotype resistance techniques (SGRT) is suboptimal. Whole blood DNA (wbDNA) from 12 HIV-1 infected patients on ART was studied by Single Genome Sequencing (SGS) and 8 of them also by Ultradeep pyrosequencing (UDP). Results were compared with contemporary and historical DRM detected in plasma by SGRT during follow up. All the contemporary DRM detected in plasma from the viremic patients were detected by SGS and UDP (20 from 7 patients and 4 from 5 patients respectively). Out of the 67 historical DRM detected in plasma and no longer present at the time of testing, 38 (57%) were detected by SGS in 12 patients and 27 out of 46 (59%) by UDP in 8 patients. Additional DRM never reported in plasma by SGRT were detected by SGS (12 from 8 patients) and UDP (10 from 8 patients). Analysis of wbDNA from HIV-1 infected patients by SGS and UDP provides proof of concept of the value of blood DNA to investigate current and archived DRM in HIV-1 infected patients on ART.

KEYWORDS:

Clonal sequencing; HIV-1; Next generation sequencing; Proviral DNA; Resistance

PMID:
29969601
DOI:
10.1016/j.jviromet.2018.06.020
[Indexed for MEDLINE]

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